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. 2020 Nov 16;117(48):30577–30588. doi: 10.1073/pnas.2013012117

Fig. 4.

Fig. 4.

The Cdc5 kinase interacts with both MutLγ and Exo1 bound on recombination sites. (A) Coimmunoprecipitation by Cdc5-TAP of Mlh1-HA, Mlh3-Myc, and Exo1-Myc from pCUP1-IME1 synchronized cells at 5.5 h in meiosis analyzed by Western blot. (B) Mlh3-Myc and Cdc5-TAP association with the indicated DSB hotspots as revealed by ChIP and qPCR at the indicated times of a pCUP1-IME1 synchronized meiotic time course. Same normalization as in Fig. 2. Values are the average of three (Mlh3-Myc) or four (Cdc5-TAP) independent experiments ± SEM (C) Coimmunoprecipitation by Cdc5-TAP of Mlh1-HA and Mlh3-Myc is independent of Exo1. Same conditions as in A. (D) Coimmunoprecipitation by Cdc5-TAP of Exo1 is independent of Exo1 interaction with MutLγ. Same conditions as in A. (E) Direct, phosphorylation-independent interaction between recombinant Exo1 and Cdc5 proteins. Western blot showing the pull-down of purified Cdc5-PM (phosphomimetic; see Materials and Methods) by Exo1-Flag, in the presence or absence of CDK1 or λ-phosphatase. See also SI Appendix, Fig. S7.